Figure 2: Rhythmicity of micturition, clock genes and Cx43 expression in WT mice is disturbed in Cry-null mice. | Nature Communications

Figure 2: Rhythmicity of micturition, clock genes and Cx43 expression in WT mice is disturbed in Cry-null mice.

From: Involvement of urinary bladder Connexin43 and the circadian clock in coordination of diurnal micturition rhythm

Figure 2

(a) A representative chart of UVVM of WT C57BL/6 mice under LD conditions followed by DD conditions. (b) Temporal UVVM every 4 h in WT mice (n=5), for 8 days under LD conditions (940 micturitions) and 5 days under DD conditions (556 micturitions). Diurnal variation of UVVM in LD conditions (F(5,20)=17.28, **P<0.005 by one-way repeated measures ANOVA) was also observed in DD conditions (F(5,20)=8.23, *P<0.05), with no significant difference among times in LD versus DD by two-way repeated measures ANOVA. (c,d) Loss of circadian rhythm of UVVM in Cry-null mice under DD conditions. Age-matched female WT, 1,493 micturitions; Cry-null, 1,009 micturitions, n=5 each. (c) A representative chart of UVVM of Cry-null mice. (d) Temporal UVVM every 4 h in Cry-null (red-diamond) and WT (black-diamond) mice. Diurnal variation detected in WT mice (F(5,20)=8.21, P<0.05 by one-way repeated measures ANOVA) was not observed in Cry-null mice. (e) Temporal Per2, Bmal1 and (f) Cx43 mRNA accumulation in the bladder in WT and Cry-null mice (n=3 for each time point). MaxCorrs of Per2, Bmal1 and Cx43 were (0.96, 0.93 and 0.85) in WT and (0.19, 0.42 and 0.38) in Cry-null mice, respectively. There was no significant difference in temporal Cx43 mRNA levels in Cry-null mice by one-way ANOVA. (g) Immunoblots showing temporal changes in protein levels of Cx43 in WT-mouse bladder (three independent samples for each time point). (h) Immunostaining of the muscle layer in mouse urinary bladder showing a difference in immunoreactivity with a decrease in Cx43 at CT4 compared with CT16. Representative photographs of three replicated experiments with similar results are shown. Bar, 50 Ī¼m. *P<0.05 and **P<0.01 by one-way ANOVA with Tukey's post hoc test in f and g. Error bars represent s.e.m. in b and d, and s.d. in eā€“g. For the relative levels, the maximal values of WT were set as 1 in e and f. F(x,y), x=DF for the time factor; y=error DF in b and c.

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